IMA FUNGUS · VOLUME 5 · no 1: 51–55 doi:10.5598/imafungus.2014.05.01.06 A new Inocyclus species (Parmulariaceae) on the neotropical fern ARTICLE Pleopeltis astrolepis Eduardo Guatimosim1, Pedro B. Schwartsburd2, and Robert W. Barreto1 1Departamento de Fitopatologia, Universidade Federal de Viçosa, CEP: 36.570-000, Viçosa, Minas Gerais, Brazil; corresponding author e-mail: [email protected] 2Departamento de Biologia Vegetal, Universidade Federal de Viçosa, CEP: 36.570-000, Viçosa, Minas Gerais, Brazil Abstract: During a survey for fungal pathogens associated with ferns in Brazil, a tar spot-causing fungus was found Key words: on fronds of Pleopeltis astrolepis. This was recognised as belonging to Inocyclus (Parmulariaceae). After comparison Ascomycota with other species in the genus, it was concluded that the fungus on P. astrolepis is a new species, described here as Brazil Inocyclus angularis sp. nov. Neotropics tropical ferns Article info: 7 January 2014; Accepted: 29 April 2014; Published: 9 May 2014. INTRODUCTION molecular-based reappraisal of the family is desirable, technical difficulties for dealing with such biotrophic The mycodiversity in Brazil is very rich, and numerous novel parasites still frustrates progress. Nevertheless the records of known and new fungal taxa have recently been description of novel taxa of Parmulariaceae should not be published, as mycological activity appears to be gaining interrupted awaiting for adequate methodologies to become momentum in this country. Poorly exploited biomes, such as available for molecular studies. Herein, a new member of the semi-arid Caatinga (Isabel et al. 2013, Leão-Ferreira et the family, found on a fern in Brazil during our ongoing al. 2013) and the savannah-like Cerrado, are having their surveys, is described based on its distinct morphology, as mycobiota surveyed and described (Hernández-Gutiérrez compared to related species. The host plant is Pleopeltis & Dianese 2014, Soares & Dianese 2014), and host-plant astrolepis, a member of a genus containing approximately focused fungal surveys (such as of native weeds and 90 species and occurring primarily in the Americas, but endangered plant species) have been conducted. Since also having species in Africa, India, and Sri Lanka (Mickel 2009, a survey of fungi occurring on ferns (also a group & Smith 2004, Otto et al. 2009, Smith & Tejero-Díez of host-plants poorly studied by mycologists) is being 2014). Pleopeltis astrolepis is a widespread fern occurring conducted in southern and south-eastern Brazil. Numerous throughout the Neotropics and extending into Mexico and mycological findings have resulted and publications are in Florida in North America (Mickel & Smith 2004). preparation to describe these. The first mycological novelty to be published as a result of this intensive study was a new genus of Parmulariaceae, Rhagadolobiopsis described on MATERIALS AND METHODS Thelypteris serrata (Thelypteridaceae) (Guatimosim et al. 2014). Parmulariaceae includes 59 genera of foliicolous Samples of leaves of the epiphytic fern Pleopeltis astrolepis biotrophic fungi, occurring mainly in the Neotropics and (Polypodiaceae) bearing minute black (tar-spot-like) colonies Paleotropics (Kirk et al. 2008). The family was recently were collected in a private garden and also on a fallen tree reviewed (Inácio & Cannon 2008). Although numerous in an Atlantic forest area in the municipality of Nova Friburgo, publications have covered this fungal family (Sivanesan state of Rio de Janeiro (Brazil), in 2013. These were dried 1970, Müller & von Arx 1973, von Arx & Müller 1975, Barr in a plant press and later examined under a dissecting 1987, Sivanesan & Hsieh 1989, Sivanesan & Sinha 1989, microscope. Freehand sections of fungal colonies on leaves Sivanesan et al. 1998, Inácio & Minter 2002a-i, Inácio were prepared and also fungal structures scraped from the 2003, Inácio & Cannon 2003a, b, Inácio 2005, Inácio et plant surface were mounted in lactophenol, lactofuchsin, al. 2011a, b, Inácio et al. 2012) it is acknowledged to be Lugol’s solution, and Melzer’s reagent. When necessary, still poorly known and many taxa in the family are still sections were made using a Microm HM 520 freezing awaiting discovery. Delimitation of existing taxa is purely microtome. Fungal structures were observed, measured morphology-based, and a serious limitation to improving (at least 30 structures), and line drawings and photographs our understanding of Parmulariaceae is the absence of any were prepared, with an Olympus BX51 light microscope fitted molecular data for fungi in the family. Although a general with a drawing tube and an Olympus E330 digital camera. © 2014 International Mycological Association You are free to share - to copy, distribute and transmit the work, under the following conditions: Attribution: You must attribute the work in the manner specified by the author or licensor (but not in any way that suggests that they endorse you or your use of the work). Non-commercial: You may not use this work for commercial purposes. No derivative works: You may not alter, transform, or build upon this work. For any reuse or distribution, you must make clear to others the license terms of this work, which can be found at http://creativecommons.org/licenses/by-nc-nd/3.0/legalcode. Any of the above conditions can be waived if you get permission from the copyright holder. Nothing in this license impairs or restricts the author’s moral rights. VOLUME 5 · NO. 1 51 Guatimosim et al. ARTICLE Fig. 1. Inocyclus angularis (VIC 39747). A–E. Colonies on P. astrolepis. F. Ascoma with asci and ascospores. h = coralloid haustoria, hy = hypostroma that connects the ascoma with the host tissue. Bars: D–E = 4 mm, F = 20 µm. Representative specimens were deposited at the herbarium Diagnosis: Differs from Inocyclus discoideus by having of the Universidade Federal de Viçosa (VIC). amphigenous ascomata and roughened versicoloured In order to observe details of ascospore germination and ascospores (i.e. with one brown and one subhyaline cell). to investigate the possibility of obtaining pure cultures of the fungus, ascospores were ejected onto the surface of PDA Type: Brazil: Rio de Janeiro: Nova Friburgo, Mury, Sítio agar in Petri plates (Crous et al. 2009). This was done by Colonial, on living leaves of Pleopeltis astrolepis, 30 Mar. attaching 1cm2 frond pieces bearing fertile ascomata to the 2013, R.W. Barreto (VIC 39747 – holotype). inside of the upper lids of Petri dishes, using vaseline, with the ascomata facing the medium. Plates were left in a growth Description: Symptoms visible as superficial amphigenous chamber adjusted to 25 ± 2 ºC under a light regime of 12 h for black tar-spot-like colonies, numerous and scattered 2 d. Additionally, ascospores were also directly ejected onto over leaves, not associated with necrosis, occasionally sterile microscope slides under similar conditions with an confluent, mostly ellipsoid to discoid, 3–10 × 7–8 mm. equivalent apparatus, but using a Petri dish lined with sterile External mycelium absent. Internal mycelium intra- and filter paper soaked with sterile water over which a sterile intercellular, deeply penetrating the mesophyll, branched, microscope slide was kept suspended on sterile glass rods. 2–3 µm, sub-hyaline, smooth. Haustoria coralloid, several per host cell, hyaline. Internal stromata absent. External stromata superficial with radiating cells, amphigenous, TAXONOMY ellipsoid to discoid, opening in circumferentially arranged locules connected to the host mesophyll at multiple points Inocyclus angularis Guatimosim & R.W. Barreto, sp. by discrete pulley wheel-like (in section) pads composed of nov. internal and external aggregations of pale brown hyphae MycoBank MB805976 connected by a peg emerging through the cuticle. Ascomata (Figs 1–2) black, initially circular, becoming ellipsoidal, producing locules arranged in one-two rings with undulated surface, Etymology: angularis, derived from the angle formed by the 800–980 × 650–670 µm, composed of dark brown textura germ-tube during ascospore germination. prismatica (cells 9 × 4 µm). In vertical section: stromata entirely superficial, strongly connected to the leaf, delimited 52 IMA FUNGUS Inocyclus angularis from a neotropical fern ARTICLE Fig. 2. Inocyclus angularis (VIC 39747). A. Ascus with hyaline ascospores (mounted on lactofuchsin). B–E. Germinated conidia, showing the nearly right angle formed by the germ tube. F. Amyloid reaction of the asci matrix on IKI. G. Detail of coralloid haustoria. Note the presence of the asexual morph (arrowed) intermixed with the sexual morph. H–I. Asexual morph with conidia. Bars: A = 10 µm, B–E = 5 µm, F = 100 µm, G, I = 20 µm, H = 50 µm. by a covering layer (above the fertile locules) and a lower cell smaller and acute, 10–13 × 3–4 µm, roughened; only layer. Covering layer 12–16.5 µm thick, black, consisting versicoloured ascospores were ejected; germination of dense dark brown-pigmented radiating cells of textura through the upper cell only, germ tubes readily folding at angularis (cells 4 × 5 µm). Lower layer underneath the approximate right angles to the main ascospore axis. hymenium adjacent to the host cuticle, 5–12 µm thick, Asexual morph intermixed with the ascomata, occupying the composed of brown to light brown textura angularis (cells same stromata located in the central region of the colonies. 2–3 × 5 µm). Locule composed of a thin basal cushion Conidia hyaline, aseptate, smooth, fusiform to clavate, with above the lower layer,